• Glutathione-S-transferase P1 isoenzyme polymorphisms, platinum-based chemotherapy, and non-small cell lung cancer.

      Booton, Richard; Ward, Timothy H; Heighway, Jim; Ashcroft, Linda; Morris, Julie; Thatcher, Nick; Christie Hospital NHS Trust, Manchester, United Kingdom. r.booten@btopenworld.com (2006-09)
      BACKGROUND: Polymorphisms within the P1 isoenzyme of GST (GSTP1) are associated with alterations in enzyme activity and may change sensitivity to platinum-based chemotherapy. We investigated the relationship between exon 5 and exon 6 GSTP1 gene polymorphisms and treatment response, hematological, and nonhematological toxicity and overall survival for patients receiving platinum-based chemotherapy for advanced non-small cell lung cancer (NSCLC). METHODS: Between 2001 and 2002, 108 patients with chemotherapy-naïve advanced NSCLC were recruited. Associations between the GSTP1 polymorphisms (Ile105Val, Thr110Ser, Ala114Val, and Asp 147Tyr) and GSTP1*A, *B, and *C haplotypes and treatment response and toxicity were evaluated using the Pearson chi and Kruskal-Wallis tests, respectively. Associations with survival were compared using Kaplan-Meier survival curves and Cox proportional hazard ratios. RESULTS: No significant associations were noted between GSTP1 polymorphisms and treatment response or survival. Significantly less neutropenic toxicity was demonstrated for patients possessing the 105Val allele (p = 0.020) or the GSTP1*B haplotype (p = 0.038). However, the variant allele GSTP1 105Val, and patients possessing a GSTP1*B allele demonstrated notable trends toward inferior response and survival. CONCLUSIONS: GSTP1 haplotype can be used to stratify hematological toxicity after platinum-based chemotherapy, but the lack of significant associations with response or survival suggests that GSTP1 polymorphisms may not be strong pharmacogenomic markers in this population. Additional large prospective studies incorporating the GSTP1 haplotype may clarify the reported discrepancies.
    • Glycolytic cancer associated fibroblasts promote breast cancer tumor growth, without a measurable increase in angiogenesis: Evidence for stromal-epithelial metabolic coupling.

      Migneco, Gemma; Whitaker-Menezes, Diana; Chiavarina, Barbara; Castello-Cros, Remedios; Pavlides, Stephanos; Pestell, Richard G; Fatatis, Alessandro; Flomenberg, Neal; Tsirigos, Aristotelis; Howell, Anthony; et al. (2010-06-06)
      Previously, we proposed a new model for understanding the Warburg effect in tumorigenesis and metastasis. In this model, the stromal fibroblasts would undergo aerobic glycolysis (a.k.a., the Warburg effect)-producing and secreting increased pyruvate/lactate that could then be used by adjacent epithelial cancer cells as "fuel" for the mitochondrial TCA cycle, oxidative phosphorylation, and ATP production. To test this model more directly, here we used a matched set of metabolically well-characterized immortalized fibroblasts that differ in a single gene. CL3 fibroblasts show a shift towards oxidative metabolism, and have an increased mitochondrial mass. In contrast, CL4 fibroblasts show a shift towards aerobic glycolysis, and have a reduced mitochondrial mass. We validated these differences in CL3 and CL4 fibroblasts by performing an unbiased proteomics analysis, showing the functional upregulation of 4 glycolytic enzymes, namely ENO1, ALDOA, LDHA and TP I1, in CL4 fibroblasts. Many of the proteins that were upregulated in CL4 fibroblasts, as seen by unbiased proteomics, were also transcriptionally upregulated in the stroma of human breast cancers, especially in the patients that were prone to metastasis. Importantly, when CL4 fibroblasts were co-injected with human breast cancer cells (MDA-MB-231) in a xenograft model, tumor growth was dramatically enhanced. CL4 fibroblasts induced a >4-fold increase in tumor mass, and a near 8-fold increase in tumor volume, without any measurable increases in tumor angiogenesis. In parallel, CL3 and CL4 fibroblasts both failed to form tumors when they were injected alone, without epithelial cancer cells. Mechanistically, under co-culture conditions, CL4 glycolytic fibroblasts increased mitochondrial activity in adjacent breast cancer cells (relative to CL3 cells), consistent with the "Reverse Warburg Effect". Notably, Western blot analysis of CL4 fibroblasts revealed a significant reduction in caveolin-1 (Cav-1) protein levels. In human breast cancer patients, a loss of stromal Cav-1 is associated with an increased risk of early tumor recurrence, metastasis, tamoxifen-resistance, and poor clinical outcome. Thus, loss of stromal Cav-1 may be an effective marker for predicting the "Reverse Warburg Effect" in the stroma of human breast cancer patients. As such, CL4 fibroblasts are a new attractive model for mimicking the "glycolytic phenotype" of cancer-associated fibroblasts. Nutrients derived from glycolytic cancer associated fibroblasts could provide an escape mechanism to confer drug-resistance during anti-angiogenic therapy, by effectively reducing the dependence of cancer cells on a vascular blood supply.
    • Glycosaminoglycans inhibit the adherence and the spreading of osteoclasts and their precursors: Role in osteoclastogenesis and bone resorption.

      Baud’huin, Marc; Ruiz-Velasco, Carmen; Jego, Gaëtan; Charrier, Céline; Gasiunas, Nijole; Gallagher, John T; Maillasson, Mike; Naggi, Annamaria; Padrines, Marc; Redini, Françoise; et al. (2011-07-20)
    • Glycosylation and epitope mapping of the 5T4 glycoprotein oncofoetal antigen.

      Shaw, David M; Woods, Andrew M; Myers, Kevin A; Westwater, Caroline; Rahi-Saund, Veena; Davies, Michael J; Renouf, David V; Hounsell, Elizabeth F; Stern, Peter L; CRC Immunology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, UK. (2002-04-01)
      The human 5T4 oncofoetal antigen is a focus for development of several antibody-directed therapies on the basis of the murine monoclonal antibody against 5T4 (mAb5T4), which recognizes a conformational epitope. 5T4 molecules are highly N-glycosylated transmembrane glycoproteins whose extracellular domain contains two regions of leucine-rich repeats (LRRs) and associated flanking regions, separated by an intervening hydrophilic sequence. Using a series of deletion and mutated cDNA constructs as well as chimaeras with the murine homologue, we have mapped the mAb5T4 epitope to the more membrane-proximal LRR2 or its flanking region. Analysis of the glycosylation of the seven consensus Asp-Xaa-Ser/Thr sites was consistent with all of the sites being glycosylated. A combination of two high-mannose chains (predominantly octasaccharide) and five mostly sialylated bi-, tri- and tetra-antennary complex chains with minor quantities of core fucose were detected. The two glycosylation sites, which are the most likely to have predominantly high-mannose chains, are in the only two regions that show significant differences between the human and the 81% identical mouse sequence. A site-directed mutation, which abolished glycosylation at one of these sites (position 192), did not alter antigenicity. The other, which is nearest to the N-terminus in the human, has an Asn-Leu-Thr to Asn-Leu-Leu conversion in the mouse, so cannot be glycosylated in the latter species. The large complex glycosylation at the other sites is likely to influence the antigenicity and tertiary structure generating the 5T4 epitope.
    • Graft dysfunction following liver transplantation: role of free radicals.

      Bzeizi, K I; Dawkes, R; Dodd, Nicholas J F; Plevris, J N; Hayes, P C; Scottish Liver Transplant Unit, Royal Infirmary of Edinburgh, UK. (1997-01)
      BACKGROUND/AIMS: Following orthotopic liver transplantation primary graft non-function occurs in about 10% of patients, and survival depends on early retransplantation. The aetiology has yet to be defined, but reperfusion injury as a result of free radical production has been considered as a possible mechanism. In this study we looked for evidence of free radical generation intraoperatively and assessed the relationship between free radical production and graft function. METHODS/RESULTS: Twenty-one patients (M:F 10:11, mean age; 53 +/- 3.8 years) who underwent liver transplantation for end-stage liver disease were studied. Free radical activity increased significantly following reperfusion, as shown by: (i) the diene conjugated method, where the percentage molar ratio increased from a baseline of 10.87 +/- 0.78% to 24.42 +/- 7.8% (p < 0.01), and (ii) by electron paramagnetic resonance, where a more than a twofold rise in radical concentration was detected (p < 0.05). The increase in free radical activity detected by the diene conjugated method was significantly higher in patients with poor outcome as compared with those who had uneventful recovery (p < 0.01). CONCLUSION: Free radical activity is increased following reperfusion of liver graft during transplantation, and the magnitude of the rise is related to the severity of graft dysfunction.
    • Granulocyte colony stimulating factor: from laboratory bench to clinical use.

      Dexter, T Michael; Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, U.K. (1994)
      A great deal has been learned about haematopoiesis since the discovery, purification and cloning of haemopoietic growth factors. The function of these growth factors is to promote survival and enhance mitosis (at higher concentrations), and influence differentiation and development of stem cells and progenitor cells. The type of mature cells produced reflects the combinations of growth factors to which the stem cells are exposed. Growth factors may also control the functional activity of mature cells. Treatment with granulocyte colony stimulating factor (G-CSF) produces neutrophils that are at least equivalent to those produced during normal haematopoiesis and does not appear to deplete the haematopoietic stem or progenitor cell populations.
    • Granulocyte-macrophage colony stimulating factor (GM-CSF) after high-dose melphalan in patients with advanced colon cancer.

      Steward, William P; Scarffe, J Howard; Dirix, L Y; Chang, James; Radford, John A; Bonnem, E; Crowther, Derek; CRC Dept of Medical Oncology, Christie Hospital, Manchester, UK. (1990-05)
      Nine patients with progressive, metastatic disease from primary carcinoma of the colon were entered into a phase I/II study using continuous intravenous infusions of granulocyte-macrophage colony-stimulating factor (GM-CSF) and high dose melphalan (120 mg m-2). GM-CSF was given alone to six patients during the first part of the study to determine a dose that would produce a peripheral leucocyte count (WCC) greater than or equal to 50 X 10(9) 1(-1) and was initially given at 3 micrograms kg-1 day-1 and escalated to 10 micrograms kg-1 day-1 after 10 days. The infusion was discontinued when the WCC exceeded 50 X 10(9) 1(-1) and after a gap of one week, melphalan was given over 30 min. GM-CSF was recommenced 8 h later and was continued until the neutrophil count had exceeded 0.5 X 10(9) 1(-1) for greater than 1 week. One patient achieved a WCC greater than 50 X 10(9) 1(-1) with GM-CSF 3 micrograms kg-1 day-1, but the other five who entered this phase of the study required dose escalation to 10 micrograms kg-1. No toxicity attributed to GM-CSF was seen. After melphalan, the median times to severe neutropenia (less than 0.5 X 10(9) 1(-1] and thrombocytopenia (greater than 20 X 10(9) 1(-1] were 6 and 9 days respectively. The median durations of neutropenia and thrombocytopenia were 14 and 10 days respectively. All patients required intensive support with a median duration of inpatient stay of 24 days. There was one treatment related death due to renal failure. One complete and two partial remissions (33% response rate) were seen but these were of short duration (median of 10 weeks). This study demonstrates that GM-CSF given by continuous intravenous infusion produces significant increments of peripheral granulocyte counts at 3 and 10 micrograms kg-1 day-1 and is not associated with any toxicity. The duration of neutropenia and thrombocytopenia induced by high-dose melphalan appears to be reduced by the subsequent administration of GM-CSF to times which are at least as short as have been reported in historical series which have used autologous bone marrow rescue.
    • Graphene Oxide promotes embryonic stem cell differentiation to haematopoietic lineage.

      Garcia-Alegria, Eva; Iluit, M; Stefanska, Monika; Silva, C; Heeg, S; Kimber, S; Kouskoff, Valerie; Lacaud, Georges; Vijayaraghavan, A; Batta, Kiran; et al. (2016)
      Pluripotent stem cells represent a promising source of differentiated tissue-specific stem and multipotent progenitor cells for regenerative medicine and drug testing. The realisation of this potential relies on the establishment of robust and reproducible protocols of differentiation. Several reports have highlighted the importance of biomaterials in assisting directed differentiation. Graphene oxide (GO) is a novel material that has attracted increasing interest in the field of biomedicine. In this study, we demonstrate that GO coated substrates significantly enhance the differentiation of mouse embryonic stem (ES) cells to both primitive and definitive haematopoietic cells. GO does not affect cell proliferation or survival of differentiated cells but rather enhances the transition of haemangioblasts to haemogenic endothelial cells, a key step during haematopoietic specification. Importantly, GO also improves, in addition to murine, human ES cell differentiation to blood cells. Taken together, our study reveals a positive role for GO in haematopoietic differentiation and suggests that further functionalization of GO could represent a valid strategy for the generation of large numbers of functional blood cells. Producing these cells would accelerate haematopoietic drug toxicity testing and treatment of patients with blood disorders or malignancies.
    • The growing burden of invasive melanoma: projections of incidence rates and numbers of new cases in six susceptible populations to 2031.

      Whiteman, D; Green, Adèle C; Olsen, C; QIMR Berghofer Medical Research Institute, 300 Herston Road, Herston, QLD 4006 Australia (2016-02-19)
      New melanoma therapies are being developed rapidly, complementing prevention and detection strategies for disease control. Estimating the future burden of melanoma is necessary for deciding how best to deploy limited resources to achieve this goal. Using three decades of cancer registry data (1982-2011) from six populations with moderate-to-high melanoma incidence (US Whites, United Kingdom (UK), Sweden, Norway, Australia, New Zealand), we applied age-period-cohort models to describe current trends and project future incidence rates and numbers of melanomas out to 2031. Between 1982-2011, melanoma rates in US Whites, UK, Sweden and Norway increased at >3% annually and are projected to continue rising until at least 2022. Melanoma incidence in Australia has been declining since 2005 (-0.7% p.a.), while melanoma incidence in New Zealand is increasing but projected to decline soon. The numbers of new melanoma cases will rise in all six populations due to aging populations and high age-specific rates in the elderly. In US Whites, annual new cases will rise from around 70,000 in 2007-11 to 116,000 in 2026-31 (79% attributable to rising age-specific rates, 21% to population growth and aging). The continued increases in case numbers in all six populations out to 2031 will increase the challenges for melanoma control.
    • Growth and development in the haemopoietic system: the role of lymphokines and their possible theirapeutic potential in disease and malignancy

      Dexter, T Michael; Moore, Michael; Departments of Experimental Haematology and Immunology, Paterson Laboratories, Christie Hospital and Holt Radium Institute, Withington, Manchester, UK. (1986)
    • Growth and differentiation in the haemopoietic system.

      Dexter, T Michael; Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, U.K. (1991-04)
    • Growth and differentiation in the hemopoietic system.

      Dexter, T Michael; Spooncer, Elaine; Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Withington, Manchester, United Kingdom. (1987)
      Hemopoiesis is regulated by a complex series of interactions, including interactions among hemopoietic cells themselves, hemopoietic cells and the extracellular matrix, hemopoietic cells and marrow stromal cells, and hemopoietic cells and growth factors. In vitro culture systems have allowed a reductionist approach to the solution of these various problems and have facilitated experiments at the mechanistic level. The hemopoietic system is organized hierarchically with multipotential self-renewing stem cells, committed progenitor cells, and mature cells. The various stimuli necessary for growth and development of these cells are rapidly being elucidated. The nature of commitment (or differentiation) remains an enigma, but model systems have been developed in which various aspects of this problem can be investigated. In this respect, growth and differentiation factors obviously have a major role to play. Now that many of these factors have been molecularly cloned (and pure target cell populations are available) their role in vivo and their mode of action can be examined.
    • Growth and differentiation of hematopoietic stem cells.

      Dexter, T Michael; Ponting, Ian L; Roberts, Ruth A; Spooncer, Elaine; Heyworth, Clare M; Gallagher, John T; Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Withington, Manchester, England. (1988)
    • Growth factor regulation of proliferation in primary cultures of small intestinal epithelium.

      Booth, Catherine; Evans, Gareth S; Potten, Christopher S; Department of Epithelial Biology, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Manchester, United Kingdom. (1995-03)
      Although the intestinal epithelium is one of the most rapidly renewing tissues, little is known about the major growth factors that control the rate of cell replacement and migration. Recently, a primary culture model has been described for the developing rat small intestinal epithelium, which permits epithelial growth while maintaining interactions with associated stromal cells, thereby possessing several contextual advantages over established cell lines (Evans et al., 1992). We have used this model to begin to determine the factors that may be involved in controlling intestinal epithelial cell proliferation. Under the conditions examined, no single growth factor promoted exclusive proliferation of epithelial cells; stromal cell proliferation was also apparent. The most potent stimulators of epithelial proliferation were insulin and insulin-like growth factor 1 (IGF-1). These factors also appeared to inhibit migration of the epithelial cells. 5-10 ng/ml EGF, 5-20 ng/ml TGF alpha, and 10-20 ng/ml PDGF also slightly increased epithelial cell numbers. Cell proliferation was inhibited by 0.1 ng/ml TGF beta-1. In Dulbecco's modified Eagle's medium (DMEM) containing 0.25 IU/ml insulin, glucose levels of 2-3 g/liter permitted epithelial growth with limited expansion of the stromal cell population. Higher levels of glucose further stimulated the nonepithelial cell types. Transferrin was also a potent stimulator of both cell types.
    • Growth factor-assisted chemotherapy--the Manchester experience.

      Crowther, Derek; Scarffe, J Howard; Howell, Anthony; Thatcher, Nick; Bronchud, M; Steward, William P; Testa, Nydia G; Dexter, T Michael; Department of Medical Oncology, Paterson Institute for Cancer Research, Christie Hospital, Manchester, UK. (1990)
      Stimulation of red cell production by erythropoietin and of granulocyte production by granulocyte colony-stimulating factor (G-CSF) and granulocyte macrophage-CSF (GM-CSF) has been demonstrated in several clinical studies. The first study to show that a human CSF could be used to shorten the period of neutropenia and reduce the risk of serious infection following intensive combination chemotherapy was carried out in Manchester using G-CSF. The period of neutropenia was significantly shortenened (by a median of 80%) and the neutrophil count levels were restored and above normal by 14 days after chemotherapy. In view of these results a further study was undertaken to examine the possibility of using intensive two weekly chemotherapy under cover of G-CSF. Treatment with Doxorubicin at doses of 75, 100, 125 and 150 mg/m2 was followed by infusion of G-CSF for 11 days. The neutrophil counts returned to normal within 12-14 days, allowing the delivery of up to three cycles of high dose chemotherapy at 14 day intervals. These studies demonstrated that intensive chemotherapy with dose-limiting myelodepression can be given with increased frequency under cover of G-CSF. Our studies using GM-CSF have also shown that administration by continuous i.v. infusion can reduce the period of life-threatening neutropenia following high dose Melphalan (120 mg/m2) without resort to autologous bone marrow transplantation (ABMT). In this study the period of granulocytopenia following Melphalan (less than 500 g x 10(9)/m2) was less than 15 days. This compares favourably with other series using high dose Melphalan followed by ABMT without CSF, where the duration of severe neutropenia was prolonged beyond three weeks. Although it appears that G-CSF and GM-CSF should be given either by continuous i.v. infusion or s.c. injection at doses between 3-10 micrograms/kg/day to obtain maximum biological effect, a great deal more work is required to determine optimum schedules and investigate the possibility of using more than one bioregulator.
    • Growth factors and the molecular control of haematopoiesis.

      Dexter, T Michael; Heyworth, Clare M; Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital, Withington, Manchester, UK. (1994)
      In the absence of appropriate growth factors, for example interleukin-3 or GM-CSF, cultured bone marrow stem cells die by a process known as apoptosis or programmed cell death. Apoptosis may occur in vivo when concentrations of specific growth factors are limiting and may be a means of regulating cell numbers. Growth factors are also essential for proliferation of bone marrow stem cells but differentiation can occur, provided there is a survival stimulus in the absence of growth factors. Combinations of growth factors may be synergistic in stimulating the survival and proliferation of multipotent stem cells. Although neither stem cell factor, nor GM-CSF alone can significantly induce the proliferation of stem cells, the combination induces the proliferation of these cells. Committed progenitor cells such as granulocyte-macrophage colony-forming cells, however, are stimulated to proliferate by GM-CSF alone, while stem cell factor in combination with GM-CSF results in only a slight additive effect. To date, most research has concentrated on the growth stimulatory factors. GM-CSF has an important role in the reversal of chemotherapy-induced myelosuppression in cancer patients and in other bone marrow disorders. A number of growth inhibitory molecules have now been identified, such as macrophage inhibitory protein-1 alpha. In the future, it is possible that improvements in cure rates may be achieved in cancer patients by combining the growth inhibitory factors with the stimulatory factors. Inhibitory factors may be given before chemotherapy to prevent toxicity and stimulatory factors may be given afterwards to treat neutropenic patients.
    • Growth factors in development, transformation, and tumorigenesis.

      Cross, Michael A; Dexter, T Michael; CRC Department of Experimental Haematology, Paterson Institute for Cancer Research, Christie Hospital and Holt Radium Institute, Manchester, England. (1991-01-25)
      Mammalian tissue development and regeneration take place within a milieu of regulatory growth factors. These affect many parameters of cell development, such that survival, proliferation, differentiation, and certain aspects of cell behavior are all influenced by a balance between stimulatory and inhibitory signals. The precise effect of any given factor is determined by the responding cell type, the concentration of factor, and the presence of other stimuli, such that some growth factors may fulfill a variety of functions under different circumstances. Classically, growth factor stimuli are transmitted into the cell via activation of specific, transmembrane receptors that modify key regulatory proteins in the cytoplasm. These in turn affect the decisions controlling proliferation and differentiation, including changes in gene expression and reactivity to other factors. There are indications that some factors may function both extra- and intracellularly and that this characteristic is correlated with potential oncogenicity. The relatively low transforming ability of extracellular factors alone is probably attributable to the limitations imposed by down-regulation of their cell surface receptors. Aberrant production of secreted growth factors can, however, play decisive roles in tumorigenesis by increasing the proliferation rate and degree of cellular autonomy and extending the area available for tumor expansion.
    • Growth factors involved in haemopoiesis.

      Dexter, T Michael; Department of Experimental Haematology, Christie Hospital & Holt Radium Institute, Withington, Manchester, UK. (1987-08)
    • Growth hormone, the insulin-like growth factor axis, insulin and cancer risk.

      Clayton, Peter E; Banerjee, I; Murray, Philip G; Renehan, Andrew G; Manchester Academic Health Sciences Centre, University of Manchester, Paediatric Endocrinology, Royal Manchester Children's Hospital, Oxford Road, Manchester, UK. peter.clayton@manchester.ac.uk (2011-01)
      Growth hormone (GH), insulin-like growth factor (IGF)-I and insulin have potent growth-promoting and anabolic actions. Their potential involvement in tumor promotion and progression has been of concern for several decades. The evidence that GH, IGF-I and insulin can promote and contribute to cancer progression comes from various sources, including transgenic and knockout mouse models and animal and human cell lines derived from cancers. Assessments of the GH-IGF axis in healthy individuals followed up to assess cancer incidence provide direct evidence of this risk; raised IGF-I levels in blood are associated with a slightly increased risk of some cancers. Studies of human diseases characterized by excess growth factor secretion or treated with growth factors have produced reassuring data, with no notable increases in de novo cancers in children treated with GH. Although follow-up for the vast majority of these children does not yet extend beyond young adulthood, a slight increase in cancers in those with long-standing excess GH secretion (as seen in patients with acromegaly) and no overall increase in cancer with insulin treatment, have been observed. Nevertheless, long-term surveillance for cancer incidence in all populations exposed to increased levels of GH is vitally important.